流苏石斛ISSR-PCR反应体系的优化  

Optimization of ISSR-PCR reaction system in Dendrobium fimbriatum Hook

作　　者：马佳梅[1] 殷寿华[1] 

机构地区：[1]中国科学院西双版纳热带植物园

出　　处：《云南大学学报（自然科学版）》2008年第S1期406-410,共5页Journal of Yunnan University(Natural Sciences Edition)

基　　金：院省合作重大专项(2000WK-07)

摘　　要：以流苏石斛为材料,对影响ISSR-PCR扩增结果的因素进行了探讨,建立了适合流苏石斛ISSR-PCR分析的最佳反应体系.在20μL反应体系中含1×buffer,2.0 mmol/L Mg2+,1.0 mmol/L dNTP s,1.0μmol/L引物,0.5 UTaqDNA聚合酶,10 ng模板DNA.扩增程序为:94℃预变性5 min;94℃变性45 s,50℃退火45 s,72℃延伸45 s,共35个循环;最后72℃延伸7 min.The major factors in the reaction system of ISSR-PCR reaction for Dendrobium fimbriatum Hook were optimized.The proper ISSR-PCR reaction system was determined as follows:20 μL reaction system containing 1×PCR buffer,2.0 mmol/L Mg2+,1.0 mmol/L dNTPs,1.0 μmol/L primer,0.5 U TaqDNA polymerase,10 ng template DNA.The amplification was after 1 cycle initial denaturation at 94 ℃ for 5 min,followed by 35 cycles of 45 s at 94 ℃ ,45 s annealing at 94 ℃,and extension of 45 s at 72 ℃,and a final 7 min extension at 72 ℃...

关 键 词：流苏石斛 ISSR 反应体系 优化 

分 类 号：S567.239[农业科学—中草药栽培]